Redox environment in stem and differentiated cells: A quantitative approach
Stem cells are believed to maintain a specific intracellular redox status through a combination of enhanced removal capacity and limited production of ROS. In the present study, we challenge this assumption by developing a quantitative approach for the analysis of the pro- and antioxidant ability of...
Main Authors: | , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Elsevier
2017-08-01
|
Series: | Redox Biology |
Online Access: | http://www.sciencedirect.com/science/article/pii/S2213231717301775 |
id |
doaj-05a41ce2b7a84595905fc35d4985e38e |
---|---|
record_format |
Article |
spelling |
doaj-05a41ce2b7a84595905fc35d4985e38e2020-11-25T01:31:28ZengElsevierRedox Biology2213-23172017-08-0112758769Redox environment in stem and differentiated cells: A quantitative approachO.G. Lyublinskaya0Ju.S. Ivanova1N.A. Pugovkina2I.V. Kozhukharova3Z.V. Kovaleva4A.N. Shatrova5N.D. Aksenov6V.V. Zenin7Yu.A. Kaulin8I.A. Gamaley9N.N. Nikolsky10Department of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, Russia; Corresponding author.Department of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, Russia; Department of Medical Physics, Institute of Physics, Nanotechnology and Telecommunications, Peter the Great St. Petersburg Polytechnic University, Polytechnicheskaya st. 29, St. Petersburg 195251, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaStem cells are believed to maintain a specific intracellular redox status through a combination of enhanced removal capacity and limited production of ROS. In the present study, we challenge this assumption by developing a quantitative approach for the analysis of the pro- and antioxidant ability of human embryonic stem cells in comparison with their differentiated descendants, as well as adult stem and non-stem cells. Our measurements showed that embryonic stem cells are characterized by low ROS level, low rate of extracellular hydrogen peroxide removal and low threshold for peroxide-induced cytotoxicity. However, biochemical normalization of these parameters to cell volume/protein leads to matching of normalized values in stem and differentiated cells and shows that tested in the present study cells (human embryonic stem cells and their fibroblast-like progenies, adult mesenchymal stem cells, lymphocytes, HeLa) maintain similar intracellular redox status. Based on these observations, we propose to use ROS concentration averaged over the cell volume instead of ROS level as a measure of intracellular redox balance. We show that attempts to use ROS level for comparative analysis of redox status of morphologically different cells could lead to false conclusions. Methods for the assessment of ROS concentration based on flow cytometry analysis with the use of H2DCFDA dye and HyPer, genetically encoded probe for hydrogen peroxide, are discussed. Keywords: Embryonic stem cells, Differentiated cells, ROS, Redox status, H2DCFDA, HyPer, Flow cytometry, Quantitative redox biologyhttp://www.sciencedirect.com/science/article/pii/S2213231717301775 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
O.G. Lyublinskaya Ju.S. Ivanova N.A. Pugovkina I.V. Kozhukharova Z.V. Kovaleva A.N. Shatrova N.D. Aksenov V.V. Zenin Yu.A. Kaulin I.A. Gamaley N.N. Nikolsky |
spellingShingle |
O.G. Lyublinskaya Ju.S. Ivanova N.A. Pugovkina I.V. Kozhukharova Z.V. Kovaleva A.N. Shatrova N.D. Aksenov V.V. Zenin Yu.A. Kaulin I.A. Gamaley N.N. Nikolsky Redox environment in stem and differentiated cells: A quantitative approach Redox Biology |
author_facet |
O.G. Lyublinskaya Ju.S. Ivanova N.A. Pugovkina I.V. Kozhukharova Z.V. Kovaleva A.N. Shatrova N.D. Aksenov V.V. Zenin Yu.A. Kaulin I.A. Gamaley N.N. Nikolsky |
author_sort |
O.G. Lyublinskaya |
title |
Redox environment in stem and differentiated cells: A quantitative approach |
title_short |
Redox environment in stem and differentiated cells: A quantitative approach |
title_full |
Redox environment in stem and differentiated cells: A quantitative approach |
title_fullStr |
Redox environment in stem and differentiated cells: A quantitative approach |
title_full_unstemmed |
Redox environment in stem and differentiated cells: A quantitative approach |
title_sort |
redox environment in stem and differentiated cells: a quantitative approach |
publisher |
Elsevier |
series |
Redox Biology |
issn |
2213-2317 |
publishDate |
2017-08-01 |
description |
Stem cells are believed to maintain a specific intracellular redox status through a combination of enhanced removal capacity and limited production of ROS. In the present study, we challenge this assumption by developing a quantitative approach for the analysis of the pro- and antioxidant ability of human embryonic stem cells in comparison with their differentiated descendants, as well as adult stem and non-stem cells. Our measurements showed that embryonic stem cells are characterized by low ROS level, low rate of extracellular hydrogen peroxide removal and low threshold for peroxide-induced cytotoxicity. However, biochemical normalization of these parameters to cell volume/protein leads to matching of normalized values in stem and differentiated cells and shows that tested in the present study cells (human embryonic stem cells and their fibroblast-like progenies, adult mesenchymal stem cells, lymphocytes, HeLa) maintain similar intracellular redox status. Based on these observations, we propose to use ROS concentration averaged over the cell volume instead of ROS level as a measure of intracellular redox balance. We show that attempts to use ROS level for comparative analysis of redox status of morphologically different cells could lead to false conclusions. Methods for the assessment of ROS concentration based on flow cytometry analysis with the use of H2DCFDA dye and HyPer, genetically encoded probe for hydrogen peroxide, are discussed. Keywords: Embryonic stem cells, Differentiated cells, ROS, Redox status, H2DCFDA, HyPer, Flow cytometry, Quantitative redox biology |
url |
http://www.sciencedirect.com/science/article/pii/S2213231717301775 |
work_keys_str_mv |
AT oglyublinskaya redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach AT jusivanova redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach AT napugovkina redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach AT ivkozhukharova redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach AT zvkovaleva redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach AT anshatrova redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach AT ndaksenov redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach AT vvzenin redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach AT yuakaulin redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach AT iagamaley redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach AT nnnikolsky redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach |
_version_ |
1725086485820997632 |